This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Olivares, I.
Right arrow Articles by Menéndez-Arias, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Olivares, I.
Right arrow Articles by Menéndez-Arias, L.

 Previous Article  |  Next Article 

Journal of Virology, August 1999, p. 6293-6298, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Second-Site Reversion of a Human Immunodeficiency Virus Type 1 Reverse Transcriptase Mutant That Restores Enzyme Function and Replication Capacity

Isabel Olivares,1 Víctor Sánchez-Merino,1 Miguel A. Martínez,2 Esteban Domingo,3 Cecilio López-Galíndez,1 and Luis Menéndez-Arias3,*

Centro Nacional de Biología Fundamental, Instituto de Salud Carlos III, 28220 Majadahonda (Madrid),1 Fundación Irsi-Caixa, Hospital Universitario Germans Trias i Pujol, Badalona (Barcelona),2 and Centro de Biología Molecular "Severo Ochoa," Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid,3 Spain

Received 17 March 1999/Accepted 10 May 1999

Nonconservative substitutions for Tyr-115 in the reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) lead to enzymes displaying lower affinity for deoxynucleoside triphosphates (dNTPs) (A. M. Martín-Hernández, E. Domingo, and L. Menéndez-Arias, EMBO J. 15:4434-4442, 1996). Several mutations at this position (Y115W, Y115L, Y115A, and Y115D) were introduced in an infectious HIV-1 clone, and the replicative capacity of the mutant viruses was monitored. Y115W was the only mutant able to replicate in MT-4 cells, albeit very poorly. Nucleotide sequence analysis of the progeny virus recovered from supernatants of four independent transfection experiments showed that the Y115W mutation was maintained. However, in all cases an additional substitution in the primer grip of the RT (M230I) emerged when the virus increased its replication capacity. Using recombinant HIV-1 RT, we demonstrate that M230I mitigates the polymerase activity defect of the Y115W mutant, by increasing the dNTP binding affinity of the enzyme. The second-site suppressor effects observed were mediated by mutations in the 66-kDa subunit of the RT, as demonstrated with chimeric heterodimers. Examination of available crystal structures of HIV-1 RT suggests a possible mechanism for restoration of enzyme activity by the second-site revertant.


* Corresponding author. Mailing address: Centro de Biología Molecular "Severo Ochoa," Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain. Phone: 34-91-3978477. Fax: 34-91-3974799. E-mail: LMENENDEZ{at}CBM.UAM.ES.


Journal of Virology, August 1999, p. 6293-6298, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Luna, E., Rodriguez-Huete, A., Rincon, V., Mateo, R., Mateu, M. G. (2009). Systematic Study of the Genetic Response of a Variable Virus to the Introduction of Deleterious Mutations in a Functional Capsid Region. J. Virol. 83: 10140-10151 [Abstract] [Full Text]  
  • Mateo, R., Mateu, M. G. (2007). Deterministic, Compensatory Mutational Events in the Capsid of Foot-and-Mouth Disease Virus in Response to the Introduction of Mutations Found in Viruses from Persistent Infections. J. Virol. 81: 1879-1887 [Abstract] [Full Text]  
  • Smith, R. A., Anderson, D. J., Preston, B. D. (2006). Hypersusceptibility to Substrate Analogs Conferred by Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase. J. Virol. 80: 7169-7178 [Abstract] [Full Text]  
  • Cases-Gonzalez, C. E., Menendez-Arias, L. (2004). Increased G->A Transition Frequencies Displayed by Primer Grip Mutants of Human Immunodeficiency Virus Type 1 Reverse Transcriptase. J. Virol. 78: 1012-1019 [Abstract] [Full Text]  
  • Delaney, W. E. IV, Yang, H., Westland, C. E., Das, K., Arnold, E., Gibbs, C. S., Miller, M. D., Xiong, S. (2003). The Hepatitis B Virus Polymerase Mutation rtV173L Is Selected during Lamivudine Therapy and Enhances Viral Replication In Vitro. J. Virol. 77: 11833-11841 [Abstract] [Full Text]  
  • Beck, J., Vogel, M., Nassal, M. (2002). dNTP versus NTP discrimination by phenylalanine 451 in duck hepatitis B virus P protein indicates a common structure of the dNTP-binding pocket with other reverse transcriptases. Nucleic Acids Res 30: 1679-1687 [Abstract] [Full Text]  
  • Gutierrez-Rivas, M., Menendez-Arias, L. (2001). A mutation in the primer grip region of HIV-1 reverse transcriptase that confers reduced fidelity of DNA synthesis. Nucleic Acids Res 29: 4963-4972 [Abstract] [Full Text]  
  • Ikuta, K., Suzuki, S., Horikoshi, H., Mukai, T., Luftig, R. B. (2000). Positive and Negative Aspects of the Human Immunodeficiency Virus Protease: Development of Inhibitors versus Its Role in AIDS Pathogenesis. Microbiol. Mol. Biol. Rev. 64: 725-745 [Abstract] [Full Text]  
  • García-Lerma, J. G., Gerrish, P. J., Wright, A. C., Qari, S. H., Heneine, W. (2000). Evidence of a Role for the Q151L Mutation and the Viral Background in Development of Multiple Dideoxynucleoside-Resistant Human Immunodeficiency Virus Type 1. J. Virol. 74: 9339-9346 [Abstract] [Full Text]  
  • Cases-Gonzalez, C. E., Gutierrez-Rivas, M., Menendez-Arias, L. (2000). Coupling Ribose Selection to Fidelity of DNA Synthesis. THE ROLE OF Tyr-115 OF HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 REVERSE TRANSCRIPTASE. J. Biol. Chem. 275: 19759-19767 [Abstract] [Full Text]